Plants were grown from germinated seeds within growth substrate up to 3–5 weeks (5–8 leaves) according to necessity, followed by stripping from growth substrate and transfer to plastic tubes containing tap-water nutrition media (pH = 7.2 ± 0.2).Upon adjustment to hydroponic media and transplant-recovery, an upper apical leaflet was submerged in-vivo within 10 ml of 1:2 diluted Fluorescein liposome solution for 96 hour period (Fluorescein – free acid, λex/λem = 496/516 nm, Sigma Aldrich, MO, USA). Secondary and tertiary roots were cut from the living root-ball at predetermined time intervals post application; they were fragmented, mounted on 1–1.2 mm thick microscopic slides, added with 50–100 µM of Propidium Iodid (depending on root thickness, λex/λem = 560/608 nm) and analyzed via Confocal Laser Scanning Microscopy, where 3D-stacks were obtained and interpreted.
Fluorescein free acid
Manufactured by Merck Group
Sourced in United States, Germany
Fluorescein (free acid) is a fluorescent dye used in various laboratory applications. It is a yellow-green crystalline compound with the chemical formula C20H12O5. Fluorescein exhibits strong green fluorescence when exposed to ultraviolet or blue light, making it a useful tool for various analytical and diagnostic purposes.
Automatically generated - may contain errors
Lab products found in correlation
Hexane, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Acetone, by Merck Group (2 mentions)
Monobasic and dibasic sodium phosphate, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
Chitosan, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Triethylamine, by Merck Group (1 mentions)
Xanthan gum, by Merck Group (1 mentions)
Sorbitol, by Merck Group (1 mentions)
3 glycidyloxypropyl trimethoxysilane, by Merck Group (1 mentions)
3 mercaptopropyl trimethoxysilane, by Merck Group (1 mentions)
Sodium benzoate, by Merck Group (1 mentions)
3 aminophenylboronic acid, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl radical dpph, by Merck Group (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid diammonium salt abts, by Merck Group (1 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
Dichloromethane, by Merck Group (1 mentions)
Rutin quercetin 3 rutinoside, by Merck Group (1 mentions)
7 protocols using fluorescein free acid
1
Fluorescein Liposome Uptake in Plant Roots
2
Silica-chitosan hybrid material synthesis
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3-Mercaptopropyltrimethoxysilane (95%), tetraethyl
orthosilicate (98%), (3-glycidyloxypropyl)trimethoxysilane (98%),
chitosan (low molecular weight), 3-aminophenylboronic acid, (3-maleimido)propyl-functionalized
silica gel, ethanol (99%), fluorescein (free acid), triethylamine
(99.7%), xanthan gum, sodium benzoate (99%), sorbitol (99%), and glycerol
were obtained from Sigma-Aldrich (UK). (3-Acryloxypropyl)trimethoxysilane
(96%) was obtained from Gelest (Morrisville, USA). Ammonium solution
(S.G. 0.88, 35%) was obtained from Fisher Scientific. Aerosil R972
Pharma was obtained from Laurence Industries (UK).
orthosilicate (98%), (3-glycidyloxypropyl)trimethoxysilane (98%),
chitosan (low molecular weight), 3-aminophenylboronic acid, (3-maleimido)propyl-functionalized
silica gel, ethanol (99%), fluorescein (free acid), triethylamine
(99.7%), xanthan gum, sodium benzoate (99%), sorbitol (99%), and glycerol
were obtained from Sigma-Aldrich (UK). (3-Acryloxypropyl)trimethoxysilane
(96%) was obtained from Gelest (Morrisville, USA). Ammonium solution
(S.G. 0.88, 35%) was obtained from Fisher Scientific. Aerosil R972
Pharma was obtained from Laurence Industries (UK).
3
Multicolor Nanoparticle Imaging Assay
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A stock solution of fluorescein-free acid (Sigma) was made up to 100 ug/mL by serial dilution in Milli-Q water. A solution containing both dye and nanoparticles was created for this experiment, formed of 50% 100 µg/mL green fluorescein stock solution, and 50% 20 nm blue FluoSpheres™ in water. 8 μL of this solution was pipetted into each inlet at the start of the experiment before starting the time-lapse acquisition (Figure 2E and F ).
The excitation wavelength for the red 10 μm polystyrene nanoparticles was 580 nm and the emission wavelength was 605 nm.
The excitation wavelength for the green fluorescein dye was 490 nm and the emission wavelength was 514 nm.
The excitation wavelength for the blue 20 nm polystyrene nanoparticles was 365 nm and the emission wavelength was 415 nm.
The excitation wavelength for the red 10 μm polystyrene nanoparticles was 580 nm and the emission wavelength was 605 nm.
The excitation wavelength for the green fluorescein dye was 490 nm and the emission wavelength was 514 nm.
The excitation wavelength for the blue 20 nm polystyrene nanoparticles was 365 nm and the emission wavelength was 415 nm.
4
Isolation and Quantification of Bioactive Compounds
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The standards 5-O-caffeoylquinic acid and rutin (quercetin-3-rutinoside) were acquired from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Cyanidin 3-O-glucoside was obtained from Polyphenols Laboratories (Sandnes, Norway). Lutein was obtained according to the method described by Meléndez-Martínez et al. [11 (link)]. Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was purchased from Fluka Chemika (Neu-Ulm, Switzerland). The reagents 2,2-diphenyl-1-picrylhydrazyl radical (DPPH∙), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS∙+), monobasic and dibasic sodium phosphate, Folin–Ciocalteu reagent, and fluorescein (free acid) were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Solvents of analytical grade ethanol, methanol, hexane, acetone, dichloromethane, acetonitrile, and formic acid were obtained from Merck (Darmstadt, Germany).
5
Thermoresponsive Polymer Synthesis
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N-isopropylacrylamide (NIPAAm), 2,2-azobisisobutyronitrile (AIBN), 1,4-dioxane, hexane, acetone, 2-(dodecylthiocarbonothioylthio)-2-methylpropionic acid, N,N′-methylenebis(acrylamide), fluorescein O-methacrylate, fluorescein (free acid), sodium thiocyanate (NaSCN), and sodium sulfate (Na2SO4) were purchased from Sigma Aldrich (St. Louis, MO, USA). Deuterated chloroform was supplied from Cambridge Isotope Ltd., (Tewksbury, MA, USA) and N,N-dimethylformamide (DMF) was purchased from Rathburn (Walkerburn, UK).
6
Isolation and Characterization of Natural Antioxidants
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The commercially available standards (+)-catechin and rutin (Quercetin-3-rutinoside) were acquired from Phytoplan GmbH (Heidelberg, Germany). Cyanidin 3-o -glucoside was purchased from Polyphenols (Sandnes, Norway). The β-carotene and β-cryptoxanthin were obtained from Sigma-Aldrich Chemie GmbH (Steinheim, Germany) and α-tocopherol was purchased from Calbiochem (Merck, Darmstadt, Germany). Violaxanthin and phytoene were isolated from natural sources by classical chromatographic techniques [10 (link)]. Luteoxanthin, neoxanthin and lutein were obtained as described by Meléndez-Martínez et al. [11 (link)].
Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was obtained from Fluka Chemika (Neu-Ulm, Switzerland). The reagents 2,2-diphenyl-1-picrylhidracyl radical (DPPH∙), monobasic and dibasic sodium phosphate, Folin Ciocalteu’s reagent and fluorescein (free acid) were purchased from Sigma–Aldrich (Steinheim, Germany). Finally, formic acid and solvents (ethanol, methanol, hexane, acetone, diclhoromethane and acetonitrile) were all of analytical grade and were obtained from Merck (Darmstadt, Germany).
Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was obtained from Fluka Chemika (Neu-Ulm, Switzerland). The reagents 2,2-diphenyl-1-picrylhidracyl radical (DPPH∙), monobasic and dibasic sodium phosphate, Folin Ciocalteu’s reagent and fluorescein (free acid) were purchased from Sigma–Aldrich (Steinheim, Germany). Finally, formic acid and solvents (ethanol, methanol, hexane, acetone, diclhoromethane and acetonitrile) were all of analytical grade and were obtained from Merck (Darmstadt, Germany).
7
Cell Culture and Imaging Protocol
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